Parathyroid hormone [1-34] improves articular cartilage surface architecture and integration and subchondral bone reconstitution in osteochondral defects in vivo

SummaryObjectiveThe 1-34 amino acid segment of the parathyroid hormone (PTH [1-34]) mediates anabolic effects in chondrocytes and osteocytes. The aim of this study was to investigate whether systemic application of PTH [1-34] improves the repair of non-osteoarthritic, focal osteochondral defects in vivo.DesignStandardized cylindrical osteochondral defects were bilaterally created in the femoral trochlea of rabbits (n = 8). Daily subcutaneous injections of 10 μg PTH [1-34]/kg were given to the treatment group (n = 4) for 6 weeks, controls (n = 4) received saline. Articular cartilage repair was evaluated by macroscopic, biochemical, histological and immunohistochemical analyses. Reconstitution of the subchondral bone was assessed by micro-computed tomography. Effects of PTH [1-34] on synovial membrane, apoptosis, and expression of the PTH receptor (PTH1R) were determined.ResultsSystemic PTH [1-34] increased PTH1R expression on both, chondrocytes and osteocytes within the repair tissue. PTH [1-34] ameliorated the macro- and microscopic aspect of the cartilaginous repair tissue. It also enhanced the thickness of the subchondral bone plate and the microarchitecture of the subarticular spongiosa within the defects. No significant correlations were established between these coexistent processes. Apoptotic levels, synovial membrane, biochemical composition of the repair tissue, and type-I/II collagen immunoreactivity remained unaffected.ConclusionsPTH [1-34] emerges as a promising agent in the treatment of focal osteochondral defects as its systemic administration simultaneously stimulates articular cartilage and subchondral bone repair. Importantly, both time-dependent mechanisms of repair did not correlate significantly at this early time point and need to be followed over prolonged observation periods

The effect of insulin-loaded chitosan particle-aggregated scaffolds in chondrogenic differentiation

Osteochondral defect repair requires a tissue engineering approach that aims at mimicking the physiological properties and structure of two different tissues (cartilage and bone) using a scaffold–cell construct. One ideal approach would be to engineer in vitro a hybrid material using a single-cell source. For that purpose, the scaffold should be able to provide the adequate biochemical cues to promote the selective but simultaneous differentiation of both tissues. In this work, attention was paid primarily to the chondrogenic differentiation by focusing on the development of polymeric systems that provide biomolecules release to induce chondrogenic differentiation. For that, different formulations of insulin-loaded chitosan particle–aggregated scaffolds were developed as a potential model system for cartilage and osteochondral tissue engineering applications using insulin as a potent bioactive substance known to induce chondrogenic differentiation. The insulin encapsulation efficiency was shown to be high with values of 70.37!0.8%, 84.26!1.76%, and 87.23!1.58% for loadings of 0.05%, 0.5%, and 5%, respectively. The in vitro release profiles were assessed in physiological conditions mimicking the cell culture procedures and quantified by Micro-BCA! protein assay. Different release profiles were obtained that showed to be dependent on the initial insulin-loading percentage. Further, the effect on prechondrogenic ATDC5 cells was investigated for periods up to 4 weeks by studying the influence of these release systems on cell morphology, DNA and glycosaminoglycan content, histology, and gene expression of collagen types I and II, Sox-9, and aggrecan assessed by real-time polymerase chain reaction. When compared with control conditions (unloaded scaffolds cultured with the standard chondrogenic-inducing medium), insulin-loaded scaffolds upregulated the Sox-9 and aggrecan expression after 4 weeks of culture. From the overall results, it is reasonable to conclude that the developed loaded scaffolds when seeded with ATDC5 can provide biochemical cues for chondrogenic differentiation. Among the tested formulations, the higher insulin-loaded system (5%) was the most effective in promoting chondrogenic differentiation.The authors would like to acknowledge the Portuguese Foundation for Science and Technology for the Ph. D. Grant to Patricia B. Malafaya (SFRH/BD/11155/2002). This work was partially supported and carried out under the scope of the European STREP Project HIPPOCRATES (NMP3-CT-2003-505758) and European NoE EXPERTISSUES (NMP3CT-2004-500283). The authors also like to acknowledge the Life and Health Sciences Research Institute (ICVS), University of Minho, for the use of their facilities, namely, to Luis Martins for histological sections slicing and H&E stain processing

Effect of transforming growth factor-ß1 (TGF-ß1) released from a scaffold on chondrogenesis in an osteochondral defect model in the rabbit

Articular cartilage repair might be stimulated by the controlled delivery of therapeutic factors. We tested the hypotheses whether TGF-ß1 can be released from a polymeric scaffold over a prolonged period of time in vitro and whether its transplantation modulates cartilage repair in vivo. Unloaded control or TGF-ß1 poly(ether-ester) copolymeric scaffolds were applied to osteochondral defects in the knee joints of rabbits. In vitro, a cumulative dose of 9 ng TGF-ß1 was released over 4 weeks. In vivo, there were no adverse effects on the synovial membrane. Defects treated with TGF-ß1 scaffolds showed no significant difference in individual parameters of chondrogenesis and in the average cartilage repair score after 3 weeks. There was a trend towards a smaller area (42.5 %) of the repair tissue that stained positive for safranin O in defects receiving TGF-ß1 scaffolds. The data indicate that TGF-ß1 is released from emulsioncoated scaffolds over a prolonged period of time in vitro and that application of these scaffolds does not significantly modulate cartilage repair after 3 weeks in vivo. Future studies need to address the importance of TGF-ß1 dose and release rate to modulate chondrogenesis

Comparison between expert listeners and continuous speech recognizers in selecting pronunciation variants.

In this paper, the performance of an automatic transcription tool is evaluated. The transcription tool is a continuous speech recognizer (CSR) which can be used to select pronunciation variants (i.e. detect insertions and deletions of phones). The performance of the CSR was compared to a reference transcription based on the judgments of expert listeners. We investigated to what extent the degree of agreement between the listeners and the CSR was affected by employing various sets of phone models (PMs). Overall, the PMs perform more similarly to the listeners when pronunciation variation is modeled. However, the various sets of PMs lead to different results for insertion and deletion processes. Furthermore, we found that to a certain degree, word error rates can be used to predict which set of PMs to use in the transcription tool

A corpus-based study of Spanish L2 mispronunciations by Japanese speakers

Abstract In a companion paper (Carranza et al.) submitted to this conference we discuss the importance of collecting specific L1-L2 speech corpora for the sake of developing effective Computer Assisted Pronunciation Training (CAPT) programs. In this paper we examine this point more deeply by reporting on a study that was aimed at compiling and analysing such a corpus to draw up an inventory of recurrent pronunciation errors to be addressed in a CAPT application that makes use of Automatic Speech Recognition (ASR). In particular we discuss some of the results obtained in the analyses of this corpus and some of the methodological issues we had to deal with. The corpus features 8.9 hours of spontaneous, semi-spontaneous and read speech recorded from 20 Japanese students of Spanish L2. The speech data was segmented and transcribed at the orthographic, canonical-phonemic and narrow-phonetic level using Praat software We report on the analyses of the combined annotations and draw up an inventory of errors that should be addressed in the training. We then consider how ASR can be employed to properly detect these errors. Furthermore, we suggest possible exercises that may be included in the training to improve the errors identified

Directions for the future of technology in pronunciation research and teaching

This paper reports on the role of technology in state-of-the-art pronunciation research and instruction, and makes concrete suggestions for future developments. The point of departure for this contribution is that the goal of second language (L2) pronunciation research and teaching should be enhanced comprehensibility and intelligibility as opposed to native-likeness. Three main areas are covered here. We begin with a presentation of advanced uses of pronunciation technology in research with a special focus on the expertise required to carry out even small-scale investigations. Next, we discuss the nature of data in pronunciation research, pointing to ways in which future work can build on advances in corpus research and crowdsourcing. Finally, we consider how these insights pave the way for researchers and developers working to create research-informed, computer-assisted pronunciation teaching resources. We conclude with predictions for future developments

Association of Hepatitis C Virus—Specific CD8+ T Cells with Viral Clearance in Acute Hepatitis C

CD8+ T lymphocytes play a major role in antiviral immune defense. Their significance for acute hepatitis C is unclear. Our aim was to correlate the CD8+ T cell response with the outcome of infection. Eighteen patients with acute hepatitis C and 19 normal donors were studied. Hepatitis C virus (HCV)—specific CD8+ T cells were identified in the enzyme-linked immunospot assay by their interferon-γ (IFN-γ) production after specific stimulation. The highest numbers of IFN-γ—producing HCV-specific CD8+ T cells were found in patients with acute hepatitis C and a self-limited course of disease during the first 6 months after onset of disease, but these numbers dropped thereafter to undetectable levels. The differences in responsiveness between patients with self-limited disease versus patients with a chronic course were statistically significant (P < .001). Our data show that the number of IFN—γ-producing HCV-specific CD8+ T cells during the first 6 months after onset of disease is associated with eradication of the HCV infectio